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Volume:4  Issue:14

July 20, 2014

Cancer Biology

Bone Resorption Assay

Authors: Carina Scholtysek
Carina ScholtysekAffiliation: Department of Internal Medicine 3, University of Erlangen-Nuremberg, Institute of Rheumatology and Immunology, Erlangen, Germany
For correspondence: carina.scholtysek@uk-erlangen.de
Bio-protocol author page: a1500
Gerhard Krönke
Gerhard KrönkeAffiliation: Department of Internal Medicine 3, University of Erlangen-Nuremberg, Institute of Rheumatology and Immunology, Erlangen, Germany
For correspondence: gerhard.kroenke@uk-erlangen.de
Bio-protocol author page: a290
 and Georg Schett
Georg SchettAffiliation: Department of Internal Medicine 3, University of Erlangen-Nuremberg, Institute of Rheumatology and Immunology, Erlangen, Germany
For correspondence: georg.schett@uk-erlangen.de
Bio-protocol author page: a1502
date: 7/20/2014, 4647 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1187 

[Abstract] The Bone resorption assay provides an easy to use protocol for quantitatively measuring in vitro osteoclast-mediated bone resorption. Osteoclasts can be seeded onto the bone slices and formation of resorption pits can be quantified via toluidinblue staining ...

Article identifier: bio-protocol.org/e1187

Developmental Biology

Hedgehog (Hh) Reporter Activity Assay

Author: Chen Zhao
Chen ZhaoAffiliation: Department of Developmental Biology, Institute for Stem Cell and Regenerative Medicine, Stanford University, Stanford, USA
For correspondence: chenzhao@stanford.edu
Bio-protocol author page: a1513
date: 7/20/2014, 4700 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1182 

[Abstract] This protocol is for testing responses of a candidate cell line/cell lines to Hh ligands or Hh pathway agonists stimulation. This protocol can also be adapted to screen small molecule libraries or biologics that contain activities to either increase ...

Article identifier: bio-protocol.org/e1182

Wnt Reporter Activity Assay

Author: Chen Zhao
Chen ZhaoAffiliation: Department of Developmental Biology, Institute for Stem Cell and Regenerative Medicine, Stanford University, Stanford, USA
For correspondence: chenzhao@stanford.edu
Bio-protocol author page: a1513
date: 7/20/2014, 8477 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1183 

[Abstract] This protocol is for testing responses of a candidate cell line/cell lines to Wnt ligands or Wnt pathway agonists stimulation. This protocol can also be adapted to screen small molecule libraries or biologics that contain activities to either increase ...

Article identifier: bio-protocol.org/e1183

Fluorescence Microscopy for Cilia in Cultured Cells and Zebrafish Embryos

Authors: Jingli Cao
Jingli CaoAffiliation: State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai, China
Bio-protocol author page: a1497
Xueliang Zhu
Xueliang Zhu Affiliation: State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai, China
Bio-protocol author page: a1498
 and Xiumin Yan
Xiumin YanAffiliation: State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai, China
For correspondence: yanx@sibcb.ac.cn
Bio-protocol author page: a1499
date: 7/20/2014, 5113 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1188 

[Abstract] Cilia are microtubule-based hair-like projections found in organisms, ranging from protozoa to mammals. This protocol provides methods for immunofluorescence staining of cilia in cultured cells and zebrafish embryos....

Article identifier: bio-protocol.org/e1188

Immunology

Isolation of ILC2 from Mouse Liver

Authors: Tamar Mchedlidze
Tamar MchedlidzeAffiliation: Medical Department 1, FAU Erlangen-Nuremberg, Erlangen, Germany
Bio-protocol author page: a1516
 and Stefan Wirtz
Stefan WirtzAffiliation: Medical Department 1, FAU Erlangen-Nuremberg, Erlangen, Germany
For correspondence: stefan.wirtz@uk-erlangen.de
Bio-protocol author page: a1517
date: 7/20/2014, 3695 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1179 

[Abstract] Group 2 innate lymphoid cells (ILC2) are a recently characterized cell population which lacks specific antigen receptors and contributes to immune responses at mucosal surfaces of lung and gut. Recently, we demonstrated that ILC2 expand in the context ...

Article identifier: bio-protocol.org/e1179

Macrophage Inflammatory Assay

Authors: Thomas J. Bartosh
Thomas J. BartoshAffiliation: Institute for Regenerative Medicine, Texas A&M University Health Science Center, Temple, USA
For correspondence: bartosh@medicine.tamhsc.edu
Bio-protocol author page: a1514
 and Joni H. Ylostalo
Joni H. YlostaloAffiliation: Institute for Regenerative Medicine, Texas A&M University Health Science Center, Temple, USA
Bio-protocol author page: a1515
date: 7/20/2014, 10537 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1180 

[Abstract] Macrophages represent a widely distributed and functionally diverse population of innate myeloid cells involved in inflammatory response to pathogens, tissue homeostasis and tissue repair (Murray and Wynn, 2011). Macrophages can be broadly grouped into ...

Article identifier: bio-protocol.org/e1180

Microbiology

In vitro EBV Infection of Mononuclear Cells that Have Been Cryo-preserved

Authors: Shanie Saghafian-Hedengren
Shanie Saghafian-HedengrenAffiliation: Department of Molecular Biosciences, Stockholm University/Wenner-Gren Institute, Stockholm, Sweden
For correspondence: shanie.hedengren@ki.se
Bio-protocol author page: a1523
Ebba Sohlberg
Ebba SohlbergAffiliation: Department of Molecular Biosciences, Stockholm University/Wenner-Gren Institute, Stockholm, Sweden
Bio-protocol author page: a1524
Jakob Theorell
Jakob TheorellAffiliation: Department of Medicine , Center for Infectious Medicine/Karolinska Institute, Stockholm, Sweden
Bio-protocol author page: a1525
Claudia Carvalho-Queiroz
Claudia Carvalho-QueirozAffiliation: Department of Molecular Biosciences, Stockholm University/The Wenner-Gren Institute, Stockholm, Sweden
Bio-protocol author page: a1526
Noémi Nagy
Noémi NagyAffiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institute, Stockholm, Sweden
Bio-protocol author page: a1527
Jan-Olov Persson
Jan-Olov PerssonAffiliation: Department of Mathematics, Stockholm University, Stockholm, Sweden
Bio-protocol author page: a1528
Caroline Nilsson
Caroline NilssonAffiliation: Department of Clinical Science and Education, Karolinska Institutet and Sachs' Children's Hospital, Stockholm, Sweden
Bio-protocol author page: a1529
Yenan T. Bryceson
Yenan T. BrycesonAffiliation: Department of Medicine, Center for Infectious Medicine/Karolinska Institute, Stockholm, Sweden
Bio-protocol author page: a1530
 and Eva Sverremark-Ekström
Eva Sverremark-EkströmAffiliation: Department of Molecular Biosciences, Stockholm University/Wenner-Gren Institute, Stockholm, Sweden
Bio-protocol author page: a1531
date: 7/20/2014, 2243 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1177 

[Abstract] Epstein-Barr Virus (EBV) is a B-lymphotropic herpesvirus which the majority of adult human population is latently-infected with. Various immunological and molecular in vitro studies have been facilitated by the use of EBV’s ability to infect and transform ...

Article identifier: bio-protocol.org/e1177

Affinofile Assay for Identifying Macrophage-Tropic HIV-1

Authors: Sarah B. Joseph
Sarah B. JosephAffiliation: Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, USA
For correspondence: sbjoseph@email.unc.edu
Bio-protocol author page: a1510
Benhur Lee
Benhur LeeAffiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, USA
Bio-protocol author page: a1511
 and Ronald Swanstrom
Ronald SwanstromAffiliation 1: Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, USA
Affiliation 2: Department of Microbiology and Immunology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, USA
For correspondence: risunc@med.unc.edu
Bio-protocol author page: a1512
date: 7/20/2014, 2502 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1184 

[Abstract] The ability to enter monocyte-derived macrophage (MDM) in vitro is commonly used to define macrophage-tropic HIV-1 despite the fact that viruses vary continuously in their ability to enter MDMs in vitro, and MDMs vary in their ability to support HIV-1 ...

Article identifier: bio-protocol.org/e1184

Chromatin Fractionation Assay in Fission Yeast

Authors: Tatsuki Kunoh
Tatsuki KunohAffiliation: Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan
For correspondence: tkunoh06@gmail.com
Bio-protocol author page: a1508
 and Toshiyuki Habu
Toshiyuki HabuAffiliation: Radiation Biology Center, Kyoto University, Kyoto, Japan
Bio-protocol author page: a1509
date: 7/20/2014, 3136 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1185 

[Abstract] The protein recruitment onto chromatin is a critical process for DNA metabolism, including DNA replication, DNA repair and DNA recombination. Especially DNA modification enzymes and checkpoint proteins are loaded onto DNA damage sites in a context-dependent ...

Article identifier: bio-protocol.org/e1185

Development of a Novel Assay for Synthesis and Hydrolysis of Sedoheptulose 1,7-bisphosphate (SBP) in vitro by Combinations of Purified Fructose 1,6-bisphosphate aldolases (FBA) Proteins and Fructose 1,6-bisphosphatases (FBPase) Proteins from Bacillus methanolicus MGA3

Authors: Jessica Stolzenberger
Jessica StolzenbergerAffiliation: Genetics of Prokaryotes, Bielefeld University, Bielefeld, Germany
Bio-protocol author page: a1503
Steffen N. Lindner
Steffen N. LindnerAffiliation: Genetics of Prokaryotes, Bielefeld University, Bielefeld, Germany
Bio-protocol author page: a1504
Marcus Persicke
Marcus PersickeAffiliation: Institute for Genome Research and Systems Biology, Bielefeld University, Bielefeld, Germany
Bio-protocol author page: a1505
Trygve Brautaset
Trygve BrautasetAffiliation: Biotechnology and Nanomedicine, SINTEF Materials and Chemistry, Trondheim, Norway
Bio-protocol author page: a1506
 and Volker F. Wendisch
Volker F. WendischAffiliation: Genetics of Prokaryotes, Bielefeld University, Bielefeld, Germany
For correspondence: volker.wendisch@uni-bielefeld.de
Bio-protocol author page: a1507
date: 7/20/2014, 2720 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1186 

[Abstract] Bacillus methanolicus (B. methanolicus) is a Gram-positive, thermotolerant, and facultative methylotrophic bacterium that can use the one-carbon (C1) compound methanol as a source of carbon and energy (Schendel et al., 1990; Arfman et al., 1997; Arfman ...

Article identifier: bio-protocol.org/e1186

Dye Release Experiments with Dextran Loaded Vesicles

Authors: Marc-Antoine Sani
Marc-Antoine SaniAffiliation: School of Chemistry, The University of Melbourne, Melbourne, Australia
For correspondence: msani@unimelb.edu.au
Bio-protocol author page: a1492
Neil M. O’Brien-Simpson
Neil M. O’Brien-SimpsonAffiliation: Melbourne Dental School, The University of Melbourne, Melbourne, Australia
Bio-protocol author page: a1488
 and Frances Separovic
Frances SeparovicAffiliation: School of Chemistry, The University of Melbourne, Melbourne, Australia
Bio-protocol author page: a1493
date: 7/20/2014, 2741 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1190 

[Abstract] Dye release experiments are a widely used method to assess the interactions between membrane-active molecules and lipid membranes. Of particular interest is the ability to assess the degree of the lipid bilayer perturbation by simultaneously encapsulating ...

Article identifier: bio-protocol.org/e1190

Bacterial Fluorescent-dextran Diffusion Assay

Authors: Neil M. O’Brien-Simpson
Neil M. O’Brien-SimpsonAffiliation: Melbourne Dental School, The University of Melbourne, Melbourne, Australia
For correspondence: neil.obs@unimelb.edu.au
Bio-protocol author page: a1488
Namfon Pantarat
Namfon PantaratAffiliation: Melbourne Dental School, The University of Melbourne, Melbourne, Australia
Bio-protocol author page: a1489
Katrina A. Walsh
Katrina A. WalshAffiliation: Melbourne Dental School, The University of Melbourne, Melbourne, Australia
Bio-protocol author page: a1490
Eric C. Reynolds
Eric C. ReynoldsAffiliation: Melbourne Dental School, The University of Melbourne, Melbourne, Australia
Bio-protocol author page: a1491
Marc-Antoine Sani
Marc-Antoine SaniAffiliation: School of Chemistry, University of Melbourne, Melbourne, Australia
Bio-protocol author page: a1492
 and Frances Separovic
Frances SeparovicAffiliation: School of Chemistry, University of Melbourne, Melbourne, Australia
Bio-protocol author page: a1493
date: 7/20/2014, 2655 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1191 

[Abstract] Antimicrobial peptides are known to disrupt bacterial membranes allowing solutes to flow across the membrane in an unregulated manner resulting in death of the organism. Disrupting the bacterial membrane would thus perturb the cells osmotic balance resulting ...

Article identifier: bio-protocol.org/e1191

Plant Science

Rice Meiotic Chromosome Spread Preparation of Pollen Mother Cells

Authors: Xingwang Li
Xingwang LiAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a1495
 and Changyin Wu
Changyin WuAffiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China
For correspondence: cywu@mail.hzau.edu.cn
Bio-protocol author page: a1496
date: 7/20/2014, 2869 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1189 

[Abstract] In this protocol, we describe a simple and efficient method for meiotic chromosome spread preparation in rice pollen mother cells. Meiotic chromosome preparation by spreading itself is an important technique for plant cytogenetics (Higgins et al., 2004; ...

Article identifier: bio-protocol.org/e1189

Stem Cell

Coculture between hMADS and Mouse Adult CM

Authors: Florence Figeac
Florence FigeacAffiliation: INSERM U955, INSERM, Créteil , France
Bio-protocol author page: a1518
Adrien Acquistapace
Adrien AcquistapaceAffiliation: INSERM U955, INSERM, Créteil, France
Bio-protocol author page: a1519
Olivier Le Coz
Olivier Le CozAffiliation: INSERM U955, INSERM, Créteil, France
Bio-protocol author page: a1520
Pierre François Lesault
Pierre François LesaultAffiliation: INSERM U955, INSERM, Créteil, France
Bio-protocol author page: a1521
 and Anne-Marie Rodriguez
Anne-Marie RodriguezAffiliation: INSERM U955, INSERM, Créteil, France
For correspondence: anne-marie.rodriguez@inserm.fr
Bio-protocol author page: a1522
date: 7/20/2014, 3345 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1178 

[Abstract] Heart failure occurring after acute myocardial infarction (MI) is among the main causes of death in western countries. Cell therapies, particularly those based on mesenchymal stem cells (MSC), represent one of the most promising approaches to repair ...

Article identifier: bio-protocol.org/e1178

Mesenchymal Stem Cell (MSC) Aggregate Formation in vivo

Authors: Thomas J. Bartosh
Thomas J. BartoshAffiliation: Institute for Regenerative Medicine, Texas A&M University Health Science Center, Temple, USA
For correspondence: bartosh@medicine.tamhsc.edu
Bio-protocol author page: a1514
 and Joni H. Ylostalo
Joni H. YlostaloAffiliation: Institute for Regenerative Medicine, Texas A&M University Health Science Center, Temple, USA
Bio-protocol author page: a1515
date: 7/20/2014, 3992 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1181 

[Abstract] Human mesenchymal stem/progenitor cells (MSCs) isolated from various adult tissues show remarkable therapeutic potential and are being employed in clinical trials for the treatment of numerous diseases (Prockop et al., 2010). While routes of cell administration ...

Article identifier: bio-protocol.org/e1181